B. Stringa, S. Garritano, A. Romanel, P. Gasperini, G. Petris, A.
Cereseto, F. Demichelis
University of Trento, Trento, Italy
Prostate cancer (PCa), the second most common cancer
among men, is a highly heritable molecularly and clinically
heterogeneous disease. In this work we focused on the study
of germline variants as putative responsible for early
somatic events in PCa. In silico analysis by our group
identified a non-coding polymorphic regulatory element at
the 7p14.3 locus that is associated with DNA repair and
hormone regulated transcript levels and with a prostate
cancer specific subclass with high genomic instability. In
vitro studies confirmed the enhancer activity of the locus
and the binding affinity for two transcription factors (TFs);
androgen receptor (AR) and CCAAT/Enhancer Binding
Protein (C/EBP) beta (CEBPB). To first proof a functional
role of the 7p14.3 locus I deleted 731bp of the genomic area
of interest in PC-3 cells by using CRISPR-Cas9. RNA-seq
analyses performed upon AR overexpression and/or
CEBPB silencing revealed significant deregulation of the
transcriptome in edited versus non-edited cells (control
cells). To well-characterize TFs binding motifs flaking the
locus, I am performing the disruption of AR and CEBPB
motifs via CRISPR-Cas9 in prostate cell lines. Moreover, to
better study the role of minor allele of the polymorphism on
genomic instability, I am currently working on the establishment of isogenic PC-3 cells via CRISPR-Cas9. Once
clones with the three possible genotypes are selected, I plan
on testing DNA damage response under relevant conditions.
This work is a proof of concept of germline predisposition
to molecularly distinct cancer subclasses and has the
potential to nominate new mechanisms of cancer
development.
Abstracts from the 51st European Society of Human Genetics Conference: Posters 459
B. Stringa: None. S. Garritano: None. A. Romanel:
None. P. Gasperini: None. G. Petris: None. A. Cereseto:
None. F. Demichelis: None.